CEFAS26-87 ITT for eDNA analysis of samples from Beyond the Reef survey

Overview

The Blue Belt Programme is the UK Government’s flagship marine conservation programme, working with a number of the UK Overseas Territories to safeguard more than four million square kilometres of the World’s oceans. Earlier this year, the Blue Belt Programme conducted its first offshore research expedition ‘Beyond the Reef’ to the Caribbean UK Overseas Territories of Anguilla, Cayman Islands and Turks and Caicos Islands. As a component of the ‘Beyond the Reef’ survey, Cefas undertook water sampling, yielding 417 filters (348 samples, 69 field negatives). Our overall aim is to identify the composition and distribution of teleost fishes and elasmobranchs around Anguilla, and the Turks and Caicos Islands in the Caribbean. We require a Supplier to process these samples using a metabarcoding workflow and provide us with the raw sequence data. As we are aiming to publish this research, we require a high level of transparency on, and input into, the laboratory procedures used. Full specification details can be found in the Bidder Pack. Services required include: - Extract DNA from the Sterivex (0.22 µm; PES) filters (348 total; triplicated per sampling station/depth; 4 L filtered total) and field negatives (69; one per station) provided by Cefas. - PCR amplify a ~170 bp fragment of the 12S rRNA region, with two primer sets: one targeting teleost fishes (MiFish-U) and another targeting elasmobranchs (MiFish-E). - Pooled PCR products will then be indexed. - Sequence the library(s) on a suitable High Throughput Sequencing Platform, e.g. NextSeq 1000 P1 XLEAP-SBS, to an appropriate level of sequence depth. - Provide Cefas with demultiplexed paired-end read sequence data.